PRK1 phosphorylates MARCKS at the PKC sites: serine 152, serine 156 and serine 163

Palmer, Ruth; Schönwasser, Dorothee C.; Rahman, Dinah; Pappin, Darryl; Herget, Thomas; Parker, Peter J.

doi:10.1016/0014-5793(95)01454-3

Cited by 24 publications

(12 citation statements)

References 22 publications

(26 reference statements)

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“…Myristoylated alanine-rich C kinase substrate (MARCKS) is a 30 kDa heat-stable member of the MARCKS family of proteins with myristoylation, MARCKS homology, and phosphorylation domains [1,2,3,4,11,12,15,19,20]. It was purified from bovine brain as a protein that underwent phosphorylation by PKC in vitro [2].…”

Section: Introductionmentioning

confidence: 99%

Development-associated myristoylated alanine-rich C kinase substrate phosphorylation in rat brain

et al. 2003

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Object: In neuronal cells, myristoylated alanine-rich C kinase substrate (MARCKS), localized to particular areas of the synaptic membrane, is active during brain development. The destination of phosphorylated MARCKS is thought to be the cytoplasm where it is probably inactive. We compared MARCKS phosphorylation in the brains of embryonic, perinatal, and adult rats to determine its possible involvement in neurogenesis. Methods: We prepared crude and partially purified extracts from various brain regions of rats aged between embryonic day 14 (E14) and 7 weeks after birth and assayed them for MARCKS phosphorylation by immunochemical methods. The isotypes of protein kinase C (PKC) were immunochemically identified in crude brain extracts from embryonic and postnatal rats. Despite negligible MARCKS phosphorylation, E16 brain extracts contained both MARCKS and PKCγ, δ, ε, and λ. MARCKS and polypeptides were clearly phosphorylated (49 and 45 kDa, respectively) in brain extracts purified on a DE52 column. Embryonic brain extracts manifested a high-molecular-weight activity capable of suppressing polypeptide phosphorylation. This activity was markedly decreased on the day of birth and almost undetectable in the brains of 9-day-old rats. Conclusions: The embryonic rat brain appears to contain a protein(s) that suppresses the phosphorylation of other proteins including MARCKS. We posit that this inhibitory activity represents a factor(s) that plays a role in the regulation of neurogenesis beginning on the day on which MARCKS appears in the embryonic brain.

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Section: Introductionmentioning

confidence: 99%

Development-associated myristoylated alanine-rich C kinase substrate phosphorylation in rat brain

et al. 2003

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“…enhanced phosphorylation of MARCKS at the PKCphosphorylated sites. Although MARCKS is a substrate for other protein kinases, such as a proline-directed kinase (Taniguchi et al, 1994) and a PKCrelated kinase (Palmer et al, 1996), there was no further phosphorylation in the presence of TPA, suggesting that the PKC substrate sites were already occupied. The PKC phosphorylation site in pp6O°" is also phosphorylated early in TPA differentiation in SH-SY5Y cells (Bjelfman et al, 1990;Pàhlman et al, 1990).…”

Section: Discussionmentioning

confidence: 99%

Effect of Continuous Phorbol Ester Treatment on Muscarinic Receptor‐Mediated Calmodulin Redistribution in SK‐N‐SH Neuroblastoma Cells

Shariat‐Madar

Goldsmith

Gnegy

1997

Journal of Neurochemistry

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Stimulation of muscarinic receptors by carbachol and activation of protein kinase C elicits the translocation of calmodulin (CaM) from membranes to cytosol in the human neuroblastoma cell line SK‐N‐SH. Our previous studies have suggested a role for protein kinase C in the regulation of CaM redistribution. To explore further the role of protein kinase C in carbachol‐induced calmodulin translocation, we treated cells for 17 h with 12‐O‐tetradecanoylphorbol 13‐acetate (TPA) to down‐regulate protein kinase C isozymes or 72 h to differentiate the cells. Treatment of SK‐N‐SH cells for 17 h with 70 nM TPA nearly abolished the effect of carbachol on CaM redistribution. After 72 h of TPA, however, the cells appeared differentiated, and the ability of carbachol to increase cytosolic CaM levels was restored. In untreated control cells, the carbachol‐mediated increase in cytosolic CaM content was mimicked by TPA and blocked by pretreatment with the selective protein kinase C inhibitor Ro 31‐8220 at 10 µM. In the 72‐h TPA‐treated cells, however, the ability of TPA to increase cytosolic CaM levels was significantly reduced, and the action of carbachol was no longer blocked by Ro 31‐8220. The effect of prolonged TPA treatment on select protein kinase C isozymes was examined by immunoblotting. Treatment of cells for either 17 or 72 h abolished the α‐isozyme in the cytosol and reduced (17 h) or abolished (72 h) the content in the membranes. In both 17‐ and 72‐h TPA‐treated cells, the ε‐isozyme was nearly abolished in the cytosol and slightly reduced in the membranes. Some protein kinase C activity may have been maintained during TPA treatment because the basal level of phosphorylation of the protein kinase C substrate myristoylated alanine‐rich C kinase substrate was enhanced in cells treated for either 17 or 72 h with TPA. The potential dissociation of carbachol and protein kinase C in eliciting increases in cytosolic CaM content was a function of prolonged TPA treatment and not differentiation per se because carbachol‐mediated increases in cytosolic CaM levels were inhibited by Ro 31‐8220 in retinoic acid‐differentiated SK‐N‐SH cells. This study demonstrates that continuous TPA treatment, although initially down‐regulating the protein kinase C‐mediated effect of carbachol on CaM redistribution, uncouples carbachol and protein kinase C at longer times.

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“…They were MARCKS (Myristoylated Alanine-Rich C Kinase Substrate) and Grin1 (G-protein-regulated inducer of neurite outgrowth). MARCKS was originally discovered as the main PKC substrate in the CNS [14], [15], but it is also phosphorylatable by proline-directed kinases [16]. Interestingly for the purpose of this study is the fact that Cdk5 phosphorylates MARCKS at Ser25 in chick neuroblasts in vivo (Ser27 in mammals) [29].…”

Section: Introductionmentioning

confidence: 91%

Searching for Novel Cdk5 Substrates in Brain by Comparative Phosphoproteomics of Wild Type and Cdk5−/− Mice

et al. 2014

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Protein phosphorylation is the most common post-translational modification that regulates several pivotal functions in cells. Cyclin-dependent kinase 5 (Cdk5) is a proline-directed serine/threonine kinase which is mostly active in the nervous system. It regulates several biological processes such as neuronal migration, cytoskeletal dynamics, axonal guidance and synaptic plasticity among others. In search for novel substrates of Cdk5 in the brain we performed quantitative phosphoproteomics analysis, isolating phosphoproteins from whole brain derived from E18.5 Cdk5+/+ and Cdk5−/− embryos, using an Immobilized Metal-Ion Affinity Chromatography (IMAC), which specifically binds to phosphorylated proteins. The isolated phosphoproteins were eluted and isotopically labeled for relative and absolute quantitation (iTRAQ) and mass spectrometry identification. We found 40 proteins that showed decreased phosphorylation at Cdk5−/− brains. In addition, out of these 40 hypophosphorylated proteins we characterized two proteins, :MARCKS (Myristoylated Alanine-Rich protein Kinase C substrate) and Grin1 (G protein regulated inducer of neurite outgrowth 1). MARCKS is known to be phosphorylated by Cdk5 in chick neural cells while Grin1 has not been reported to be phosphorylated by Cdk5. When these proteins were overexpressed in N2A neuroblastoma cell line along with p35, serine phosphorylation in their Cdk5 motifs was found to be increased. In contrast, treatments with roscovitine, the Cdk5 inhibitor, resulted in an opposite effect on serine phosphorylation in N2A cells and primary hippocampal neurons transfected with MARCKS. In summary, the results presented here identify Grin 1 as novel Cdk5 substrate and confirm previously identified MARCKS as a a bona fide Cdk5 substrate.

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PRK1 phosphorylates MARCKS at the PKC sites: serine 152, serine 156 and serine 163

Cited by 24 publications

References 22 publications

Development-associated myristoylated alanine-rich C kinase substrate phosphorylation in rat brain

Development-associated myristoylated alanine-rich C kinase substrate phosphorylation in rat brain

Effect of Continuous Phorbol Ester Treatment on Muscarinic Receptor‐Mediated Calmodulin Redistribution in SK‐N‐SH Neuroblastoma Cells

Searching for Novel Cdk5 Substrates in Brain by Comparative Phosphoproteomics of Wild Type and Cdk5−/− Mice

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